WH2 and proline-rich domains of WASP-family proteins collaborate to accelerate actin filament elongation.

WASP-family proteins are known to promote assembly of branched actin networks by stimulating the filament-nucleating activity of the Arp2/3 complex. Here, we show that WASP-family proteins also function as polymerases that accelerate elongation of uncapped actin filaments. When clustered on a surface, WASP-family proteins can drive branched actin networks to grow much faster than they could by direct incorporation of soluble monomers. This polymerase activity arises from the coordinated action of two regulatory sequences: (i) a WASP homology 2 (WH2) domain that binds actin, and (ii) a proline-rich sequence that binds profilin-actin complexes. In the absence of profilin, WH2 domains are sufficient to accelerate filament elongation, but in the presence of profilin, proline-rich sequences are required to support polymerase activity by (i) bringing polymerization-competent actin monomers in proximity to growing filament ends, and (ii) promoting shuttling of actin monomers from profilin-actin complexes onto nearby WH2 domains. Unoccupied WH2 domains transiently associate with free filament ends, preventing their growth and dynamically tethering the branched actin network to the WASP-family proteins that create it. Collaboration between WH2 and proline-rich sequences thus strikes a balance between filament growth and tethering. Our work expands the number of critical roles that WASP-family proteins play in the assembly of branched actin networks to at least three: (i) promoting dendritic nucleation; (ii) linking actin networks to membranes; and (iii) accelerating filament elongation

Poleward transport of Eg5 by dynein–dynactin in Xenopus laevis egg extract spindles

Molecular motors are required for spindle assembly and maintenance during cell division. How motors move and interact inside spindles is unknown. Using photoactivation and photobleaching, we measure mitotic motor movement inside a dynamic spindle. We find that dynein–dynactin transports the essential motor Eg5 toward the spindle poles in Xenopus laevis egg extract spindles, revealing a direct interplay between two motors of opposite directionality. This transport occurs throughout the spindle except at the very spindle center and at the spindle poles, where Eg5 remains stationary. The variation of Eg5 dynamics with its position in the spindle is indicative of position-dependent functions of this motor protein. Our results suggest that Eg5 drives microtubule flux by antiparallel microtubule sliding in the spindle center, whereas the dynein-dependent concentration of Eg5 outside the spindle center could contribute to parallel microtubule cross-linking. These results emphasize the importance of spatially differentiated functions of motor proteins and contribute to our understanding of spindle organization

CLIP-170 tracks growing microtubule ends by dynamically recognizing composite EB1/tubulin-binding sites

The microtubule cytoskeleton is crucial for the internal organization of eukaryotic cells. Several microtubule-associated proteins link microtubules to subcellular structures. A subclass of these proteins, the plus end–binding proteins (+TIPs), selectively binds to the growing plus ends of microtubules. Here, we reconstitute a vertebrate plus end tracking system composed of the most prominent +TIPs, end-binding protein 1 (EB1) and CLIP-170, in vitro and dissect their end-tracking mechanism. We find that EB1 autonomously recognizes specific binding sites present at growing microtubule ends. In contrast, CLIP-170 does not end-track by itself but requires EB1. CLIP-170 recognizes and turns over rapidly on composite binding sites constituted by end-accumulated EB1 and tyrosinated α-tubulin. In contrast to its fission yeast orthologue Tip1, dynamic end tracking of CLIP-170 does not require the activity of a molecular motor. Our results demonstrate evolutionary diversity of the plus end recognition mechanism of CLIP-170 family members, whereas the autonomous end-tracking mechanism of EB family members is conserved

Changes in self-schema structure in cognitive therapy for major depressive disorder: a randomized clinical trial.

Negative cognitive structure (particularly for interpersonal content) has been shown in some research to persist past a current episode of depression and potentially to be a stable marker of vulnerability for depression (D. J. A. Dozois, 2007; D. J. A. Dozois & K. S. Dobson, 2001a). Given that cognitive therapy (CT) is highly effective for treating the acute phase of a depressive episode and that this treatment also reduces the risk of relapse and recurrence, it is possible that CT may alter these stable cognitive structures. In the current study, patients were randomly assigned to CT+ pharmacotherapy (n = 21) or to pharmacotherapy alone (n = 21). Both groups evidenced significant and similar reductions in level of depression (as measured with the Beck Depression Inventory-II and the Hamilton Rating Scale for Depression), as well as automatic thoughts and dysfunctional attitudes. However, group differences were found on cognitive organization in favor of individuals who received the combination of CT+ pharmacotherapy. The implications of these results for understanding mechanisms of change in therapy and the prophylactic nature of CT are discussed

Changes in Core Beliefs (Early Maladaptive Schemas) and Self-Representation in Cognitive Therapy and Pharmacotherapy for Depression

Randomized clinical trials suggest that cognitive therapy (CT) is comparable to antidepressant medication for the acute treatment of depression. Compelling data also indicate that CT has an added prophylactic benefit relative to pharmacotherapy (PT). The purpose of this study was to examine cognitive change in CT for depression. Participants (N = 42) met diagnostic criteria for a current major depressive episode and were randomly assigned to CT + PT or PT. Participants completed indices of depressive symptomatology, core beliefs (i.e., early maladaptive schemas), and self-attribute redundancy before and after therapy. Self-attribute redundancy was conceptualized as a form of schema organization and operationalized as the number of similar traits that permeate different aspects of self (e.g., as a partner, friend, employee). Treatment change was evident in both groups on self-reported core belief domains, with few between-group differences. Although no group differences were found on attribute redundancy at pre-treatment, there was a significant increase in positive redundancy at post-treatment favoring CT + PT. No group differences were found for negative content. These findings suggest that something about CT may uniquely impact self-representation and that CT may operate by bolstering compensatory schemas

Modeling the mental health service utilization decisions of university undergraduates: A discrete choice conjoint experiment

Objective: We modeled design factors influencing the intent to use a university mental health service. Participants: Between November 2012 and October 2014, 909 undergraduates participated. Method: Using a discrete choice experiment, participants chose between hypothetical campus mental health services. Results: Latent class analysis identified three segments. A Psychological/Psychiatric Service segment (45.5%) was most likely to contact campus health services delivered by psychologists or psychiatrists. An Alternative Service segment (39.3%) preferred to talk to peer-counselors who had experienced mental health problems. A Hesitant segment (15.2%) reported greater distress but seemed less intent on seeking help. They preferred services delivered by psychologists or psychiatrists. Simulations predicted that, rather than waiting for standard counseling, the Alternative Service segment would prefer immediate access to E-Mental health. The Usual Care and Hesitant segments would wait 6 months for standard counseling. Conclusions: E-Mental Health options could engage students who may not wait for standard services.This project was supported by the Jack Laidlaw Chair in Patient-Centered Health Care and a grant from the Canadian Health Services Research Foundation